Chemical compound and its use

ABSTRACT

The present application relates to a novel tetrahydroquinoline derivative, to a process for its preparation, to its use on its own or in combination for treating and/or preventing diseases and to its use for preparing medicaments, in particular as an inhibitor of the cholesterol ester transfer protein (CETP) for the treatment and/or prevention of cardiovascular disorders, in particular hypolipoproteinaemias, dyslipidaemias, hypertriglyceridaemias, hyperlipidaemias, hypercholesterolaemias and arteriosclerosis.

The present application relates to a novel tetrahydroquinolinederivative, to a process for this preparation, to its use on its own orin combination for treating and/or preventing diseases and to its usefor preparing medicaments, in particular as an inhibitor of thecholesterol ester transfer protein (CETP) for the treatment and/orprevention of cardiovascular disorders, in particularhypolipoproteinaemias, dyslipidaemias, hypertriglyceridaemias,hyperlipidaemias, hypercholesterolaemias and arteriosclerosis.

Coronary heart disease caused by arteriosclerosis is one of the maincauses of death in modern society. In a large number of studies, it wasshown that low plasma concentrations of HDL cholesterol are an importantrisk factor for the development of arteriosclerosis [Barter and Rye,Atherosclerosis 121, 1-12 (1996)]. HDL (high density lipoprotein), inaddition to LDL (low density lipoprotein) and VLDL (very low densitylipoprotein), is a class of lipoproteins whose most important functionis the transport of lipids, such as, for example, cholesterol,cholesterol esters, triglycerides, fatty acids or phospholipids, in theblood. High LDL cholesterol concentrations (>160 mg/dl) and low HDLcholesterol concentrations (<40 mg/dl) contribute substantially to thedevelopment of arteriosclerosis [ATP III Guidelines, Report of the NCEPExpert Panel]. In addition to coronary heart disease, unfavourableHDL/LDL ratios also promote the development of peripheral vasculardisorders and stroke. Accordingly, novel methods for elevating HDLcholesterol in the plasma are a therapeutically useful advance in theprevention and treatment of arteriosclerosis and the disordersassociated therewith.

Cholesterol ester transfer protein (CETP) mediates the exchange ofcholesterol esters and triglycerides between the different lipoproteinsin the blood [Tall, J. Lipid Res. 34, 1255-74 (1993)]. Of particularimportance here is the transfer of cholesterol esters from HDL to LDL,which results in a reduction of the plasma HDL cholesterolconcentration. Accordingly, inhibition of CETP should result in elevatedplasma HDL cholesterol concentrations and a reduction of the plasma LDLcholesterol concentrations and thus in a therapeutically useful effecton the lipid profile in the plasma [McCarthy, Medicinal Res. Rev. 13,139-59 (1993); Sitori, Pharmac. Ther. 67, 443-47 (1995); Swenson, J.Biol. Chem. 264, 14318 (1989)]. Tetrahydroquinolines havingpharmacological activity are known from EP-A-818 448, WO 99/14215, WO99/15504 and WO 03/028727. Substituted tetrahydronaphthalenes havingpharmacological activity are known from WO 99/14174.

It is an object of the present invention to provide novel substances forcontrolling disorders, in particular cardiovascular disorders, whichsubstances have an improved therapeutic profile.

The present invention provides the compounds of the structural formula(I)

in which R represents cyclopentyl or isopropyl, and their salts,solvates and solvates of the salts.

The present invention provides in particular the compound having thesystematic name(5S)-4-cyclohexyl-2-cyclopentyl-3-{(S)-fluoro[4-(trifluoromethyl)phenyl]methyl}-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-5-oland the structural formula (Ia)

and its salts, solvates and solvates of the salts.

The present invention in particular also provides the compound havingthe systematic name(5S)-4-cyclopentyl-3-{(S)-fluoro[4-(trifluoromethyl)phenyl]methyl}-2-isopropyl-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-5-oland the structural formula (Ib)

and its salts, solvates and solvates of the salts.

Hereinbelow, the compounds of the formula (I), (Ia) and (Ib) arereferred to in the singular as “compound according to the invention”;however, the description relates to both compounds.

The compound according to the invention can also be present in otherstereoisomeric forms (enantiomers, diastereomers). The present inventioncomprises all enantiomers, diastereomers and their respective mixtures.From such mixtures of enantiomers and/or diastereomers, thestereoisomerically uniform components can be isolated in a known manner.Preferred is the S-configuration at C-5 and at C-3′ shown in formula(I).

In the context of the present invention, preferred salts arephysiologically acceptable salts of the compound according to theinvention. However, salts which for their part are unsuitable forpharmaceutical applications but which can be used, for example, forisolating or purifying the compound according to the invention are alsoincluded.

Physiologically acceptable salts of the compound according to theinvention include acid addition salts of mineral acids, carboxylic acidsand sulphonic acids, for example salts of hydrochloric acid, hydrobromicacid, sulphuric acid, phosphoric acid, methanesulphonic acid,ethanesulphonic acid, toluenesulphonic acid, benzenesulphonic acid,naphthalenedisulphonic acid, acetic acid, trifluoroacetic acid,propionic acid, lactic acid, tartaric acid, malic acid, citric acid,fumaric acid, maleic acid and benzoic acid.

Physiologically acceptable salts of the compound according to theinvention also include salts of customary bases, such as, by way ofexample and by way of preference, alkali metal salts (for example sodiumsalts and potassium salts), alkaline earth metal salts (for examplecalcium salts and magnesium salts) and ammonium salts, derived fromammonia or organic amines having 1 to 16 carbon atoms, such as, by wayof example and by way of preference, ethylamine, diethylamine,triethylamine, ethyldiisopropylamine, monoethanolamine, diethanolamine,triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine,dibenzylamine, N-methylmorpholine, arginine, lysine, ethylenediamine andN-methylpiperidine.

In the context of the invention, solvates refer to those forms of thecompound according to the invention which, in solid or liquid state,form a complex by coordination with solvent molecules. Hydrates are aspecial form of solvates where the coordination is with water. In thecontext of the present invention, preferred solvates are hydrates.

Moreover, the present invention also includes prodrugs of the compoundaccording to the invention. The term “prodrugs” includes compounds whichfor their part may be biologically active or inactive but are converted(for example metabolically or hydrolytically) into the compoundaccording to the invention during their residence time in the body.

In the context of the invention, (C₁-C₄) alkyl represents astraight-chain or branched alkyl radical having 1 to 4 carbon atoms. Thefollowing radicals may be mentioned by way of example and by way ofpreference: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl and tert-butyl.

The invention also provides a process for preparing the compound of theformula (Ia) according to the invention, characterized in that thecompound of the formula (IIa)

is initially, by asymmetric reduction, converted into the compound ofthe formula (IIIa)

which is then either

-   [A] by introduction of a hydroxyl protective group reacted to give a    compound of the formula (IVa)

-   -   in which    -   PG represents a hydroxyl protective group, preferably a radical        of the formula —SiR¹R²R³, in which    -    R¹, R² and R³ are identical or different and represent (C₁-C₄)        alkyl,    -   and then, by diastereoselective reduction, converted into a        compound of the formula (Va)

-   -   in which PG is as defined above,        or in the reverse order of the reaction sequence

-   [B] initially reduced diastereoselectively to give the compound of    the formula (VIa)

-    which is then, by regioselective introduction of the hydroxyl    protective group PG, converted into a compound of the formula (Va),    the compound of the formula (Va) is then, using a fluorinating    agent, reacted to give a compound of the formula (VIIa)

in which PG is as defined above,and the hydroxyl protective group PG is then cleaved off by customarymethods giving the compound of the formula (Ia)and the compound of the formula (Ia) is, if appropriate, converted withthe appropriate (i) solvents and/or (ii) bases and/or acids into itssolvates, salts and/or solvates of the salts.

The compound of the formula (IIa) can be prepared by reacting thecompounds of the formulae (VIII), (IX) and (Xa)

in a 3-component reaction in the presence of a protic acid or Lewis acidwith one another to give the compound of the formula (XIa)

and then oxidizing this compound to the compound of the formula (IIa).

The invention furthermore provides a process for preparing the compoundof the formula (Ib) according to the invention, characterized in thatthe compound of the formula (IIb)

is initially, by asymmetric reduction, converted into the compound ofthe formula (IIIb)

which is then either

-   [A] by introduction of a hydroxyl protective group reacted to give a    compound of the formula (IVb)

-   -   in which    -   PG represents a hydroxyl protective group, preferably a radical        of the formula —SiR¹R²R³, in which    -    R¹, R² and R³ are identical or different and represent (C₁-C₄)        alkyl,    -   and then, by diastereoselective reduction, converted into a        compound of the formula (Vb)

-   -   in which PG is as defined above,        or in the reverse order of the reaction sequence

-   [B] initially reduced diastereoselectively to give the compound of    the formula (VIb)

-    which is then, by regioselective introduction of the hydroxyl    protective group PG, converted into a compound of the formula (Vb),    the compound of the formula (Vb) is then, using a fluorinating    agent, reacted to give a compound of the formula (VIIb)

in which PG is as defined above,and the hydroxyl protective group PG is then cleaved off by customarymethods giving the compound of the formula (Ib)and the compound of the formula (Ib) is, if appropriate, converted withthe appropriate (i) solvents and/or (ii) bases or acids into itssolvates, salts and/or solvates of the salts.

The compound of the formula (IIb) can be prepared by reacting thecompounds of the formulae (VIII), (IX) and (Xb)

in a 3-component reaction in the presence of a protic acid or Lewis acidwith one another to give the compound of the formula (XIb)

and then oxidizing this compound to give the compound of the formula(IIb).

Compounds of the formulae (VIII), (IX) and (Xa) and (Xb) arecommercially obtainable, known from the literature or can be preparedanalogously to processes known from the literature (cf. also WO 99/14215and WO 03/028727).

Suitable inert solvents for the individual process steps are, forexample, ethers, such as diethyl ether, diisopropyl ether, dioxane,tetrahydrofuran, glycol dimethyl ether or diethylene glycol dimethylether, hydrocarbons, such as benzene, toluene, xylene, hexane,cyclohexane or mineral oil fractions, or halogenated hydrocarbons, suchas dichloromethane, trichloromethane, carbon tetrachloride,1,2-dichloroethane, trichloroethylene or chlorobenzene. It is alsopossible to use mixtures of the solvents mentioned.

The reductions in process steps (II)→(III), (IV)→(V) and (III)→(VI) aregenerally carried out using reducing agents suitable for reducingketones to hydroxyl compounds. These include, in particular, complexaluminium hydrides or borohydrides, such as, for example, lithiumhydride, sodium hydride, potassium hydride, zinc borohydride, lithiumaluminium hydride, diisobutylaluminium hydride (DIBAH), sodiumbis-(2-methoxyethoxy)aluminium dihydride, lithium trialkylborohydridesor lithium trialkoxyaluminium hydrides, or borane complexes, such as,for example, borane tetrahydrofuran, borane dimethyl sulphide or boraneN,N-diethylaniline complex.

The asymmetric reduction in process step (II)→(III) is carried out inthe presence of catalytic amounts (0.01 to 0.3 mol equivalents) ofenantiomerically pure (1R,2S)-1-aminoindan-2-ol as chiral inductor. Thereducing agent which is preferably used for this purpose is boraneN,N-diethylaniline complex. The reaction is generally carried out in oneof the ethers listed above or in toluene, preferably in tetrahydrofuran,in a temperature range of from −80° C. to +50° C., preferably from 0° C.to +30° C.

The reducing agent used for the reductions (IV)→(V) and (III)→(VI) ispreferably lithium aluminium hydride or DIBAH. The reactions aregenerally carried out in one of the ethers listed above or in toluene,preferably in tetrahydrofuran or toluene, in a temperature range of from−80° C. to +50° C., in the case of lithium aluminium hydride preferablyfrom 0° C. to +30° C. and in the case of DIBAH preferably from −80° C.to +30° C.

A preferred hydroxyl protective group for process steps (III)→(IV) or(VI)→(V) is a silyl group, such as, for example, trimethylsilyl,triethylsilyl, triisopropylsilyl or tert-butyldimethylsilyl. Particularpreference is given to tert-butyldimethylsilyl. The silyl group isgenerally introduced in one of the abovementioned hydrocarbons,halogenated hydrocarbons, ethers or in dimethylformamide as solvent, inthe presence of a base, such as, for example, triethylamine,N,N-diisopropylethylamine, pyridine, 2,6-lutidine or4-N,N-dimethylaminopyridine (DMAP).

In process step (III)→(IV), the silylating agent used is preferablytert-butyldimethylsilyl trifluoromethanesulphonate in combination with2,6-lutidine as base. The reaction is preferably carried out indichloromethane or toluene, in a temperature range of from −40° C. to+40° C., preferably from −20° C. to +30° C.

In process step (VI)→(V), the silylating agent used is preferablytert-butyldimethylsilyl chloride in combination with triethylamine andDMAP as bases. The reaction is preferably carried out indimethylformamide, in a temperature range of from 0° C. to +100° C.,preferably from +20° C. to +80° C.

The fluorination in process step (VI)→(VII) is generally carried out inone of the abovementioned hydrocarbons or halogenated hydrocarbons or inacetonitrile, preferably in toluene or dichloromethane, usingdiethylaminosulphur trifluoride (DAST) or morpholino-sulphur trifluorideas fluorinating agent. The reaction is generally carried out in atemperature range of from −80° C. to +40° C., preferably from −60° C. to+20° C.

Removal of a silyl protective group in process step (VII)→(I) isgenerally carried out with the aid of acids, such as, for example,hydrochloric acid or trifluoroacetic acid, or with the aid of fluorides,such as, for example, hydrogen fluoride or tetrabutylammonium fluoride(TBAF). Suitable inert solvents are the abovementioned ethers, alcohols,such as methanol or ethanol, or mixtures of the solvents mentioned. Theremoval is preferably carried out using TBAF in tetrahydrofuran assolvent. The reaction is generally carried out in a temperature range offrom −20° C. to +60° C., preferably from 0° C. to +30° C.

The condensation reaction (VIII)+(IX)+(X)→(XI) is generally carried outin one of the abovementioned ethers, in alcohols, such as methanol,ethanol, n-propanol or isopropanol, in acetonitrile or in mixtures ofthe solvents mentioned. Preference is given to using diisopropyl ether.

Protic acids suitable for this process step are, in general, organicacids, such as, for example, acetic acid, trifluoroacetic acid, oxalicacid or para-toluenesulphonic acid, or inorganic acids, such as, forexample, hydrochloric acid, sulphuric acid, or phosphoric acid. Alsosuitable are Lewis acids, such as, for example, aluminium chloride orzinc chloride. Preference is given to trifluoroacetic acid.

In general, the reaction is carried out in a temperature range of from0° C. to +120° C., preferably from +20° C. to +80° C.

The oxidation (dehydrogenation) in process step (XI)→(II) is generallycarried out in one of the halogenated hydrocarbons listed above, or, ifappropriate, in alcohols, such as methanol or ethanol, in acetonitrileor in water. Suitable oxidizing agents are, for example, nitric acid,cerium (IV) ammonium nitrate, 2,3-dichloro-5,6-dicyano-1,4-benzoquinone(DDQ), pyridinium chlorochromate (PCC), osmium tetroxide, manganesedioxide or a catalytic dehydrogenation using platinum dioxide orpalladium-on-carbon. Preference is given to an oxidation using DDQ indichloromethane as solvent. The oxidation is generally carried out in atemperature range of from −50° C. to +100° C., preferably from 0° C. to+40° C.

The individual process steps can be carried out at atmospheric, elevatedor reduced pressure (for example from 0.5 to 5 bar). In general, theprocess steps are carried out at atmospheric pressure.

The preparation of the compound according to the invention can beillustrated by the synthesis scheme below:

The compound according to the invention has an unforeseeable usefulpharmacological acitivity spectrum. Accordingly, it is suitable for useas a medicinally active compound for the treatment and/or prophylaxis ofdiseases in humans and animals.

The compound according to the invention opens up a further treatmentalternative and represents an advance of pharmacy. In comparison to theknown and previously employed preparations, the compound according tothe invention shows an improved spectrum of action.

It is preferably distinguished by great specificity, good tolerabilityand fewer side-effects, and also a reduced toxicity, in particular inthe cardiovascular area and in the liver area.

An advantage of the compound according to the invention is its highactivity in human plasma. A further advantage of the compound accordingto the invention is a reduced potential for interactions withmetabolizing enzymes, in particular the cytochrome P450 enzymes andespecially the cytochrome P450 3A4 enzyme. In addition, the compoundaccording to the invention has a reduced tendency to deposit itself infatty tissues.

The compound of the formula (I) according to the invention has usefulpharmacological properties and can be used for the prevention andtreatment of disorders. The compound according to the invention is inparticular a highly effective inhibitor of the cholesterol estertransfer protein (CETP) and stimulates reverse cholesterol transport. Itelevates the HDL cholesterol concentration in the blood. The compoundaccording to the invention is particularly suitable for the treatmentand for primary or secondary prevention of coronary heart disease, forexample myocardial infarction. In addition, the compound according tothe invention can be used for the treatment and prevention ofarteriosclerosis, restenosis, strokes and Alzheimer's disease. Moreover,the compound according to the invention can also be used for thetreatment and prevention of hypolipoproteinaemias, dyslipidaemias,hypertriglyceridaemias, hyperlipidaemias, hypercholesterolaemias,adiposity, obesity, pancreatitis, insulin-dependent andnon-insulin-dependent diabetes, diabetic sequelae such as, for example,retinopathy, nephropathy and neuropathy, of combined hyperlipidaemiasand of the metabolic syndrome.

The pharmacological action of the compound according to the inventioncan be determined using the CETP inhibition tests described below.

The present invention furthermore provides the use of the compoundaccording to the invention for the treatment and/or prevention ofdisorders, in particular the disorders mentioned above.

The present invention furthermore provides the use of the compoundaccording to the invention for preparing a medicament for the treatmentand/or prevention of disorders, in particular the disorders mentionedabove.

The present invention furthermore provides a method for the treatmentand/or prevention to of disorders, in particular the disorders mentionedabove, using an effective amount of the compound according to theinvention.

The present invention furthermore provides medicaments comprising thecompound according to the invention and one or more further activecompounds, for the treatment and/or prevention of disorders. Activecompounds suitable for combinations are, by way of example and by way ofpreference:

-   -   antidiabetics,    -   substances having antithrombotic action,    -   hypotensive substances,    -   lipid metabolism-modifying substances,    -   anti-inflammatory substances,    -   substances which stabilize arteriosclerotic plaque.

The compound of the formula (I) according to the invention canpreferably be combined with one or more

-   -   antidiabetics mentioned in the Roten Liste [red list] 2002/II,        chapter 12,    -   agents having antithrombotic action, by way of example and by        way of preference from the group of the platelet aggregation        inhibitors or the anticoagulants,    -   hypotensive agents, by way of example and by way of preference        from the group of the calcium antagonists, angiotensin AII        antagonists, ACE inhibitors beta blockers, phosphodiesterase        inhibitors, stimulators of soluble guanylate cyclase, cGMP        enhancers and diuretics, and/or    -   active compounds which modify lipid metabolism, by way of        example and by way of preference from the group of the thyroid        receptor agonists, the cholesterol synthase inhibitors, such as        HMG-CoA reductase inhibitors, squalene synthase inhibitors,        squalene epoxidase inhibitors or oxidosqualene cyclase        inhibitors, the ACAT inhibitors, MTP inhibitors, PPAR agonists,        fibrates, lipase inhibitors, cholesterol absorption inhibitors,        bile acid reabsorption inhibitors, polymeric bile acid adsorbers        and the lipoprotein(a) antagonists.

Antidiabetics are to be understood as meaning, by way of example and byway of preference, insulin and insulin derivatives, and also orallyeffective compounds with hypoglycaemic action.

Here, insulin and insulin derivatives include both insulins of animal,human or biotechnological origin and mixtures thereof.

The orally effective compounds with hypoglycaemic action include, by wayof example and by way of preference, sulphonylureas, biguanidines,meglitinide derivatives, oxadiazolidinones, thiazolidinediones,glucosidase inhibitors, glucagon antagonists, GLP-1 agonists, insulinsensitizers, inhibitors of liver enzymes involved in the stimulation ofgluconeogenesis and/or glycogenolysis, modulators of glucose uptake andpotassium channel openers, such as, for example, those disclosed in WO97/26265 and WO 99/03861.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with insulin.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a sulphonylurea, such as, by wayof example and by way of preference, tolbutamide, glibenclamide,glimepiride, glipizide or gliclazide.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a biguanide, such as, by way ofexample and by way of preference, metformin.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a meglitinide derivative, suchas, by way of example and by way of preference, repaglinide ornateglinide.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a PPARgamma agonist, for examplefrom the class of the thiazolidinediones, such as, by way of example andby way of preference, pioglitazone or rosiglitazone.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a mixed PPARalpha/gamma agonist,such as, by way of example and by way of preference, GI-262570(farglitazar), GW 2331, GW 409544, AVE 8042, AVE 8134, AVE 0847, MK-0767(KRP-297) or AZ-242.

Agents with antithrombotic action are to be understood as meaning,preferably, compounds from the group of the platelet aggregationinhibitors, such as, by way of example and by way of preference,aspirin, clopidogrel, ticlopidine or dipyridamole, or of theanticoagulants.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a thrombin inhibitor, such as,by way of example and by way of preference, ximelagatran, melagatran,bivalirudin or clexane.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a GPIIb/IIIa antagonist, suchas, by way of example and by way of preference, tirofiban or abciximab.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a factor Xa inhibitor, such as,by way of example and by way of preference, DX 9065a, DPC 906, JTV 803or BAY 59-7939.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with heparin or alow-molecular-weight (LMW) heparin derivative.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a vitamin K antagonist, such as,by way of example and by way of preference, coumarin.

Hypotensive agents are to be understood as meaning, by way of exampleand by way of preference, compounds from the group of the calciumantagonists, such as, by way of example and by way of preference, thecompounds nifedipine, amlodipine, nitrendipine, nisoldipine, verapamilor diltiazem, of the angiotensin AII antagonists, ACE inhibitors, betablockers and the diuretics.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with an antagonist of the alpha 1receptors.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with reserpine, minoxidil, diazoxide,dihydralazine, hydralazine and nitrous oxide-releasing substances, suchas, by way of example and by way of preference, glycerol nitrate orsodium nitroprusside.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with an angiotensin AII antagonist,such as, by way of example and by way of preference, losartan,valsartan, candesartan, telmisartan, embusartan, irbesartan, olmesartan,tasosartan or saprisartan.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with an ACE inhibitor, such as, byway of example and by way of preference, enalapril, captopril, ramipril,delapril, fosinopril, quinopril, perindopril or trandolapril.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a beta blocker, such as, by wayof example and by way of preference, propranolol or atenolol.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a diuretic, such as, by way ofexample and by way of preference, furosemide.

Lipid metabolism-modifying agents are to be understood as meaning, byway of example and by way of preference, compounds from the group of thethyroid receptor agonists, the cholesterol synthesis inhibitors, such asHMG-CoA reductase inhibitors or squalene synthesis inhibitors, the ACATinhibitors, MTP inhibitors, PPAR agonists, fibrates, cholesterolabsorption inhibitors, bile acid reabsorption inhibitors, lipaseinhibitors, polymeric bile acid adsorbers and the lipoprotein(a)antagonists.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a thyroid receptor agonist, suchas, by way of example and by way of preference, D-thyroxine,3,5,3′-triiodothyronine (T3), CGS 23425 or axitirome (CGS 26214).

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a squalene synthesis inhibitor,such as, by way of example and by way of preference, BMS-188494 or TAK475.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with an ACAT inhibitor, such as, byway of example and by way of preference, avasimibe, eflucimibe orCS-505.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a cholesterol absorptioninhibitor, such as, by way of example and by way of preference,ezetimibe, tiqueside or pamaqueside.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a bile acid reabsorbtioninhibitor, such as, by way of example and by way of preference,barixibat, AZD 7508, SC 435, SC 635, S-8921, 264W94 or HM 1453.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with an MTP inhibitor, such as, byway of example and by way of preference, implitapide, BMS-201038 orR-103757.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a PPARalpha agonist, such as,for example, the fibrates fenofibrate, clofibrate, bezafibrate,ciprofibrate or gemfibrozil, or such as, by way of example and by way ofpreference, GW 9578, GW 7647, LY-518674 or NS-220.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a PPARdelta agonist, such as, byway of example and by way of preference, GW 501516.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a mixed PPARalpha/gamma agonist,such as, by way of example and by way of preference, GI-262570(farglitazar), GW 2331, GW 409544, AVE 8042, AVE 8134, AVE 0847, MK-0767(KRP-297) or AZ-242.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a mixed PPARalpha/gamma/deltaagonist, such as, by way of example and by way of preference, MCC-555.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a lipase inhibitor from thegroup of the endothelial lipase inhibitors, the pancreatic lipaseinhibitors, the gastric lipase inhibitors, the hormone-sensitive lipaseinhibitors or the hepatic lipase inhibitors.

In a particularly preferred embodiment of the invention, the compound ofthe formula (I) is administered in combination with an inhibitor ofpancreatic lipase, preferably from the class of the lipstatins, such as,by way of example, orlistat.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a polymeric bile acid adsorber,such as, by way of example and by way of preference, cholestyramine,colestipol, colesolvam, CholestaGel or colestimide.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with a lipoprotein(a) antagonist,such as, by way of example and by way of preference, gemcabene calcium(CI-1027) or nicotinic acid.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with an antagonist of the niacinreceptor, such as, by way of example and by way of preference, niaspan,acipimox or niceritrol.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with an antioxidant, such as, by wayof example and by way of preference, probucol, AGI1067 or Bo 653.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with an LDL receptor inducer, suchas, by way of example, lifibrol.

In a preferred embodiment of the invention, the compound of the formula(I) is administered in combination with an HMG-CoA reductase inhibitorfrom the class of the statins, such as, by way of example and by way ofpreference, lovastatin, simvastatin, pravastatin, fluvastatin,atorvastatin, rosuvastatin, cerivastatin or pitavastatin.

The present invention also provides combinations of the compound of theformula (I) with substances which reduce the gene expression of HMG-CoAreductase. Such substances may, for example, be inhibitors of HMG-CoAreductase transcription or HMG-CoA reductase translation Inhibition ofHMG-CoA reductase gene expression may be effected, for example, byinhibiting S1P (Site-1) protease, or by lowering the SREBP (sterolreceptor binding protein) concentration.

The present invention also provides combinations of the compound of theformula (I) with substances which may have anti-inflammatory actionand/or stabilize arteriosclerotic plaque. Such substances may, forexample, be active compounds from the class of the NSAIDs, the PAF-AHantagonists or the chemokine receptor antagonists, such as, by way ofexample, IL-8 receptor antagonists or MCP-1 antagonists.

The active compound combinations according to the invention have usefulpharmacological properties and can be used for the prophylaxis andtreatment of disorders. The active compound combinations according tothe invention are particularly suitable for the treatment and for theprimary or secondary prevention of coronary heart disease, for exampleof miocardial infarction. Additionally, they can be used for thetreatment and prevention of arteriosclerosis, restenosis, stroke andAlzheimer's disease. In addition, the active compound combinationsmentioned can also be employed for the treatment and prevention ofhypolipoproteinaemias, dyslipidaemias, hypertriglyceridaemias,hyperlipidaemias, hypercholesterolaemias, adiposity, obesity,pancreatitis, insulin-dependent and non-insulin-dependent diabetes,diabetic sequelae, such as, for example, retinopathy, nephropathy andneuropathy, of combined hyperlipidemias and of the metabolic syndrome.Furthermore, the active compound combinations according to the inventionare suitable for treating hypertension, heart failure, angina pectoris,ischaemias and inflammatory disorders.

The present invention furthermore provides medicaments comprising thecompound according to the invention, usually together with one or moreinert non-toxic pharmaceutically suitable auxiliaries, and their use forthe purposes mentioned above.

The compound according to the invention can act systemically and/orlocally. For this purpose, it can be administered in the suitablemanner, such as, for example, orally, parenterally, pulmonarily,nasally, sublingually, lingually, buccally, rectally, dermally,transdermally, conjunctivally, otically or as an implant or stent.

For these administration routes, the compound according to the inventioncan be administered in suitable administration forms.

Suitable for oral administration are administration forms which workaccording to the prior art, deliver the compound according to theinvention rapidly and/or in modified form and which comprise thecompound according to the invention in crystalline and/or amorphisizedand/or dissolved form, such as, for example, tablets (uncoated or coatedtablets, for example tablets provided with enteric coatings or coatingswhich dissolve in a delayed manner or are insoluble and which controlthe release of the compound according to the invention), tablets whichrapidly disintegrate in the oral cavity or films/wafers,films/lyophilizates, capsules (for example hard or soft gelatincapsules), sugar-coated tablets, granules, pellets, powders, emulsions,suspensions, aerosols or solutions.

Parenteral administration can be carried out with avoidance of anabsorption step (for example intravenously, intraarterially,intracardially, intraspinally or intralumbally) or with involvement ofan absorption (for example intramuscularly, subcutaneously,intracutaneously, percutaneously or intraperitoneally). Suitableadministration forms for parenteral administration are, inter alia,injection and infusion preparations in the form of solutions,suspensions, emulsions, lyophilizates or sterile powders.

Suitable for the other administration routes are, for examplepharmaceutical forms for inhalation (inter alia powder inhalers,nebulizers), nasal drops, solutions or sprays, tablets to beadministered lingually, sublingually or bucally, films/wafers orcapsules, suppositories, aural and ophthalmic preparations, vaginalcapsules, aqueous suspensions (lotions, shaker mixtures), lipophilicsuspensions, ointments, creams, transdermal therapeutic systems (forexample patches), milk, pastes, foams, dusting powders, implants orstents.

Preference is given to oral or parenteral administration, in particularto oral administration.

The compound according to the invention can be converted into theadministration forms mentioned. This may take place in a manner knownper se by mixing with inert non-toxic pharmaceutically suitableauxiliaries. These auxiliaries include, inter alia, carriers (forexample microcrystalline cellulose, lactose, mannitol), solvents (forexample liquid polyethylene glycols), emulsifiers and dispersants orwetting agents (for example sodium dodecylsulphate, polyoxysorbitanoleate), binders (for example polyvinylpyrrolidone), synthetic andnatural polymers (for example albumin), stabilizers (for exampleantioxidants, such as, for example, ascorbic acid), colorants (forexample inorganic pigments, such as, for example, iron oxides) and tasteand/or odour correctants.

In general, it has been found to be advantageous to administer, in thecase of parenteral administration, amounts of from about 0.001 to 1mg/kg, preferably about 0.01 to 0.5 mg/kg, of body weight to obtaineffective results. In the case of oral administration, the dosage isfrom about 0.01 to 100 mg/kg, preferably about 0.01 to 20 mg/kg and veryparticularly preferably 0.1 to 10 mg/kg, of body weight.

In spite of this, it may, if appropriate, be necessary to depart fromthe amounts mentioned, namely depending on the body weight, theadministration route, the individual response to the active compound,the type of preparation and the time or interval at which administrationtakes place. Thus, in some cases, it may be sufficient to manage withless than the abovementioned minimum amount, while in other cases theupper limit mentioned has to be exceeded. In the case of theadministration of relatively large amounts, it may be advisable todivide these into a number of individual doses over the course of theday.

The following exemplary embodiments illustrate the invention. Theinvention is not limited to the examples.

The percentages in the tests and examples below are, unless indicatedotherwise, percentages by weight; parts are parts by weight. Solventratios, dilution ratios and stated concentrations of liquid/liquidsolutions are in each case based on volume.

A. EXAMPLES Abbreviations and Acronyms

CE Cholesterol esterCETP Cholesterol ester transfer proteinDAST Dimethylaminosulphur trifluorideDCI Direct chemical ionization (in MS)DDQ 2,3-Dichloro-5,6-dicyano-1,4-benzoquinonede Diastereomeric excess

DMF N,N-dimethylformamide

DMSO Dimethyl sulphoxideEDTA Ethylenediamine-N,N,N′,N′-tetraacetic acidee Enantiomeric excesseq. Equivalent(s)ESI Electrospray ionization (in MS)

h Hour(s)

HDL High density lipoproteinHPLC High pressure, high performance liquid chromatographyLC/MS Liquid chromatography-coupled mass spectroscopyLDL Low density lipoprotein

min Minute(s)

MS Mass spectroscopyNMR Nuclear magnetic resonance spectroscopyR_(t) Retention time (in HPLC)SPA Scintillation proximity assayTBAF Tetrabutylammonium fluorideTBDMSOTf tert-Butyldimethylsilyl trifluoromethanesulphonateTFA Trifluoroacetic acid

THF Tetrahydrofuran

HPLC and LC/MS methods:

Method 1: Column: Chiralpak IA, 250 mm×4.6 mm; mobilephase:isohexane/1-propanol 97:3; flow rate: 1.0 ml/min; UV detection:254 nm.

Method 2: Instrument: HP 1100 with DAD detection; column: KromasilRP-18, 60 mm×2 mm, 3.5 μm; mobile phase A: 5 ml of HClO₄/l of water,mobile phase B: acetonitrile; gradient: 0 min 2% B→0.5 min 2% B→4.5 min90% B→9 min 90% B; flow rate: 0.75 ml/min; temperature: 30° C.; UVdetection: 210 nm.

Method 3 (LC/MS): MS instrument: MicromaB ZQ; HPLC instrument: HP 1100Series; UV DAD; column: Phenomenex Synergi 2μ. Hydro-RP Mercury 20 mm×4mm; mobile phase A: 11 of water+0.5 ml of 50% strength formic acid,mobile phase B: 11 of acetonitrile+0.5 ml of 50% strength formic acid;gradient: 0.0 min 90% A→2.5 min 30% A→3.0 min 5% A→4.5 min 5% A; flowrate: 0.0 min→1 ml/min 2.5 min/3.0 min/4.5 min 2 ml/min; oven: 50° C.;UV detection: 210 nm.

Starting Materials and Intermediates: (a) Example 1A2-cyclopentyl-4-cyclohexyl-7,7-dimethyl-3-(4-trifluoromethylbenzoyl)-4,6,7,8-tetrahydro-1H-quinolin-5-one

15.0 g (53 mmol, 1.2 eq.) of3-amino-3-cyclopentyl-1-(4-trifluoromethylphenyl)propenone (preparationaccording to WO 03/028727, example 4) are initially charged in 500 ml ofdiisopropyl ether, and 6.80 ml (88 mmol, 2.0 eq.) of trifluoroaceticacid and 6.19 g (44 mmol, 1 eq.) of 5,5-dimethylcyclohexane-1,3-dioneare added. After 10 min of stirring at room temperature, 7.1 ml (66mmol, 1.5 eq.) of cyclohexanecarbaldehyde are added. The mixture is thenheated under reflux on a water separator for 15 h. After cooling, themixture is stirred in an ice bath for 30 min. The resulting precipitateis filtered off with suction and washed with cold diisopropyl ether.

Yield: 3.13 g (14% of theory)

¹H-NMR (CDCl₃, 300 MHz): δ=0.77-2.05 (m, 20H), 1.17 (s, 6H), 2.21 (m,2H), 2.40 (2d, 2H), 3.48 (sept, 1H), 3.79 (d, 1H), 5.85 (s, 1H), 7.66(d, 2H), 7.78 (d, 2H) ppm.

MS (ESIpos): m/z=500 [M+H]⁺.

Example 2A2-cyclopentyl-4-cyclohexyl-7,7-dimethyl-3-(4-trifluoromethylbenzoyl)-7,8-dihydro-6H-quinolin-5-one

3.13 g (6.3 mmol) of the compound from Example 1A are dissolved in 64 mlof dichloromethane, and 1.42 g (6.3 mmol) of2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) are added a little at atime at 0° C. With stirring, the mixture is warmed to room temperatureover a period of 3 h. The mixture is concentrated on a rotary evaporatorand the residue is purified by chromatography (silica gel, mobilephase:cyclohexane/ethyl acetate 5:1).

Yield: 3.07 g (98.6% of theory)

¹H-NMR (CDCl₃, 400 MHz): δ=1.01-1.22 (m, 2H), 1.10 (s, 3H), 1.18 (s,3H), 1.35-2.00 (m, 16H), 2.50-2.69 (m, 3H), 3.07 (s, 2H), 3.35 (m, 1H),7.75 (d, 2H), 7.94 (m, 2H) ppm.

MS (ESIpos): m/z=498 [M+H]⁺.

Example 3A[(5S)-2-cyclopentyl-4-cyclohexyl-5-hydroxy-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-3-yl](4-trifluoromethylphenyl)methanone

178 mg (1.2 mmol, 0.08 eq.) of (1R,2S)-1-aminoindan-2-ol are initiallycharged in 400 ml of THF, and 9.73 g (60 mmol, 4 eq.) of boraneN,N-diethylaniline complex are added at room temperature. After theevolution of gas has ceased, the mixture is cooled to 0° C. and 7.42 g(14.9 mmol, 1 eq.) of the compound from Example 2A, dissolved in 400 mlof THF, are added. With stirring, the mixture is allowed to warm to roomtemperature over a period of 16 h. After the reaction has ended, 20 mlof methanol are added to the reaction mixture, the mixture isconcentrated and the residue is purified by chromatography (silica gel,mobile phase:isohexane/ethyl acetate gradient).

Yield: 6.97 g (93.5% of theory)

According to method 1, the enantiomeric excess is determined as 97.6%ee.

¹H-NMR (CDCl₃, 400 MHz): δ=0.90-2.10 (m, 27H), 2.55 (m, 1H), 2.70 (m,1H), 2.80-3.40 (m, 2H), 5.18 (br. s, 1H), 6.8 (m, 2H), 7.40-8.40 (br. m,4H) ppm.

MS (DCI/NH₃): m/z=500 [M+H]⁺.

Example 4A((5S)-5-{[tert-butyl(dimethyl)silyl]oxy}-4-cyclohexyl-2-cyclopentyl-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-3-yl)[4-(trifluoromethyl)phenyl]methanone

Under argon, 6.0 g (12.0 mmol) of the compound from Example 3A areinitially charged in 45 ml of dry toluene. At room temperature, 5.15 g(48.0 mmol, 4 eq.) of 2,6-lutidine are then added, and the mixture iscooled to −16° C. 6.35 g (24.0 mmol, 2 eq.) of tert-butyldimethylsilyltrifluoromethanesulphonate in 15 ml of toluene are added dropwise tothis solution. After 15 min, the reaction mixture is warmed to 0° C. andstirred at this temperature for 80 min. For work-up, 0.1 N hydrochloricacid (186 ml) is added and the mixture is, after warming to roomtemperature, extracted with ethyl acetate. The aqueous phase isre-extracted three times with ethyl acetate, the combined organic phasesare washed with saturated sodium bicarbonate solution and with saturatedsodium chloride solution and this aqueous phase for its part isre-extracted with ethyl acetate. The combined organic phases are driedover sodium sulphate, filtered and concentrated under reduced pressure,and the residue is purified by chromatography (silica gel, mobilephase:isohexane/ethyl acetate gradient).

Yield: 5.96 g (80.8% of theory)

¹H-NMR (CDCl₃, 300 MHz): δ=0.13 (s, 3H), 0.19 (s, 3H), 0.86 (s, 9H),0.99-2.08 (m, 26H), 2.43-2.69 (m, 2H), 2.92-3.29 (m, 2H), 5.24 (br. s,1H), 6.8 (m, 2H), 7.48-8.03 (br. m, 4H) ppm.

MS (DCI/NH₃): m/z=614 [M+H]⁺.

Example 5A(S)-((5S)-5-{[tert-butyl(dimethyl)silyl]oxy}-4-cyclohexyl-2-cyclopentyl-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-3-yl)[4-(trifluoromethyl)phenyl]methanol

At 0° C., 10.25 ml of a 1 M solution of lithium aluminium hydride (10.25mmol, 1.1 eq.) in THF are added dropwise to a solution of 5.72 g (9.3mmol) of the compound from Example 4A in 116 ml of dry THF. Withstiffing, the mixture is warmed to room temperature over a period of 6h. For work-up, 120 ml of a saturated sodium potassium tartrate solutionare added carefully. After the evolution of gas has ceased, the mixtureis extracted three times with ethyl acetate, the combined organic phasesare washed with a 1:1 mixture of sodium bicarbonate solution andsaturated sodium chloride solution and this aqueous phase for its partis re-extracted with ethyl acetate. The combined organic phases aredried over sodium sulphate, filtered and concentrated under reducedpressure. The residue is purified chromatographically and at the sametime separated into the diastereomers of the product (column: ChiralpakAD, 500 mm×40 mm, 20 μm; mobile phase:isohexane/isopropanol 97.5:2.5;flow rate: 50 ml/min).

Yield: 2.5 g (43.6% of theory)

Diastereomeric purity: 96.9% de, R_(t)=4.15 min (method 1).

¹H-NMR (CDCl₃, 400 MHz): δ=0.18 (br. s, 6H), 0.90 (s, 9H), 1.03-2.09 (m,26H), 2.10-2.33 (br. m, 1H), 2.37-3.06 (m, 3H), 3.33 (m, 1H), 5.24 and5.47 (2 br. s, 1H), 6.49 and 6.64 (2 br. s, 1H), 7.31-7.64 (m, 4H) ppm.

MS (ESIpos): m/z=616 [M+H]⁺.

Syn Diastereomer(R)-((5S)-5-{[tert-butyl(dimethyl)silyl]oxy}-4-cyclohexyl-2-cyclopentyl-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-3-yl)[4-(trifluoromethyl)phenyl]methanol

Yield: 3.56 g (61.2% of theory)

Diastereomeric purity: 98.6% de, R_(t)=2.77 min (method 1).

Example 6A(5S)-5-{[tert-butyl(dimethyl)silyl]oxy}-4-cyclohexyl-2-cyclopentyl-3-{(S)-fluoro[4-(trifluoromethyl)phenyl]methyl}-7,7-dimethyl-5,6,7,8-tetrahydroquinoline

At −55° C. and under argon, 3.21 ml of diethylaminosulphur trifluoride(24.3 mmol, 1.5 eq.) are added dropwise to a solution of 9.96 g (16.2mmol) of the compound from Example 5A in 300 ml of dry toluene. Themixture is stirred at this temperature for 1 h and then at roomtemperature for a further 2 h. For work-up, 120 ml of a saturated sodiumbicarbonate solution are added carefully. The mixture is extracted threetimes in total with ethyl acetate. The combined organic phases arewashed with saturated sodium chloride solution, dried over sodiumsulphate, filtered and concentrated under reduced pressure. The crudeproduct is purified by filtration through silica gel (mobilephase:cyclohexane/ethyl acetate 9:1).

Yield: 9.93 g (99.3% of theory)

¹H-NMR (CDCl₃, 400 MHz): δ=0.20 (br. s, 6H), 0.91 (s, 9H), 1.03-2.14 (m,26H), 2.42-3.04 (m, 3H), 3.35 (m, 1H), 5.26 and 5.52 (2 br. s, 1H),7.10-7.50 (m, 3H), 7.62 (d, 2H) ppm.

MS (ESIpos): m/z=618 [M+H]⁺.

WORKING EXAMPLES: (a) Example 1(5S)-4-cyclohexyl-2-cyclopentyl-3-{(S)-fluoro[4-(trifluoromethyl)phenyl]methyl}-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-5-ol

At 0° C., 40.1 ml of a 1 M solution of tetrabutylammonium fluoride (40.1mmol, 2.5 eq.) in THF are added dropwise to a solution of 9.91 g (16.0mmol) of the compound from Example 6A in 200 ml of dry THF. The mixtureis warmed to room temperature and stirred at this temperature for 16 h.For work-up, the mixture is diluted with 100 ml of ethyl acetate andwashed twice with in each case 100 ml of water and with 50 ml ofsaturated sodium chloride solution. The organic phase is dried oversodium sulphate, filtered and concentrated under reduced pressure. Thecrude product is purified chromatographically (silica gel, mobilephase:isohexane/ethyl acetate 100:0→1:1).

Yield: 7.7 g (95.3% of theory)

¹H-NMR (CDCl₃, 300 MHz): δ=1.02 (s, 3H), 1.17 (s, 3H), 1.08-2.15 (m,21H), 2.59-3.00 (m, 3H), 3.51 (m, 1H), 5.13 (m, 1H), 7.34 (d, 2H), 7.39(d, 1H), 7.61 (d, 2H) ppm.

MS (DCI): m/z=504 [M+H]⁺

HPLC (method 2): R_(t)=5.20 min.

B. ASSESSMENT OF THE PHARMACOLOGICAL ACTIVITY (a) B-I. CETP-InhibitionTesting B-I.1. Obtainment of CETP

CETP is obtained in partially purified form from human plasma bydifferential centrifugation and column chromatography and used for thetest. To this end, human plasma is adjusted to a density of 1.21 g perml using NaBr and centrifuged at 4° C. at 50 000 rpm for 18 h. Thebottom fraction (d >1.21 g/ml) is applied to a Sephadex®Phenyl-Sepharose 4B (Pharmacia) column, washed with 0.15 M NaCl/0.001 Mtris-HCl pH 7.4 and then eluted with distilled water. The CETP-activefractions are pooled, dialysed against 50 mM sodium acetate pH 4.5 andapplied to a CM-Sepharose® column (Pharmacia). The mixture is theneluted using a linear gradient (0-1 M NaCl). The pooled CETP fractionsare dialysed against 10 mM tris/HCl pH 7.4 and then further purified bychromatography on a Mono Q® column (Pharmacia).

B-I.2. CETP Fluorescence Test

Measurement of the CETP-catalysed transfer of a fluorescent cholesterolester between liposomes [modified according to the procedure of Bisgaieret al., J. Lipid Res. 34, 1625 (1993)]:

For the production of the donor liposomes, 1 mg of cholesteryl4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-dodecanoate(cholesteryl BODIPY® FL C₁₂, Molecular Probes) is dissolved in 600 μl ofdioxane with 5.35 mg of triolein and 6.67 mg of phosphatidylcholine withgentle warming in an ultrasonic bath and this solution is added veryslowly with ultrasonication to 63 ml of 50 mM tris/HCl, 150 mM NaCl, 2mM EDTA buffer pH 7.3 at room temperature. The suspension is thenultrasonicated under an N₂ atmosphere for 30 minutes in the Bransonultrasonic bath at about 50 watts, the temperature being kept at about20° C.

The acceptor liposomes are obtained analogously from 86 mg ofcholesteryl oleate, 20 mg of triolein and 100 mg of phosphatidylcholinedissolved in 1.2 ml of dioxane and 114 ml of the above buffer byultrasonication at 50 watts (20° C.) for 30 minutes.

B-I.2.1. CETP Fluorescence Test with Enriched CETP

For testing, a test mix consisting of 1 part of above buffer, 1 part ofdonor liposomes and 2 parts of acceptor liposomes is used.

50 μl of test mix are treated with 48 μl of enriched CETP fraction (1-3μg), obtained from human plasma by means of hydrophobic chromatography,and 2 μl of a solution of the substance to be investigated in DMSO andincubated at 37° C. for 4 hours.

The change in the fluorescence at 485/535 nm is a measure of the CEtransfer; the inhibition of the transfer in comparison to the controlbatch without substance is to determined

IC₅₀ [nM] Example fluorescence No. test 1 25B-I.2.2. CETP Fluorescence Test with Human Plasma

6 μl (12% v/v) of donor liposomes and 1 μl (2% v/v) of a solution of thesubstance to be investigated in DMSO are added to 42 μl (86% v/v) ofhuman plasma (Sigma P9523), and the mixture is incubated at 37° C. for24 h.

The change in the fluorescence at 510/520 nm (gap width 2.5 nm) is ameasure of the CE transfer; the inhibition of the transfer in comparisonto the control batch without substance is determined

IC₅₀ [nM] Example fluorescence test in No. human plasma 1 50

B-I.2.3. Ex Vivo-CETP Fluorescence Test

10 μl of buffer and 2 μl of serum are added to 80 μl of test mix, andthe mixture is incubated at 37° C. for 4 h.

The change in the fluorescence at 485/535 nm is a measure for the CEtransfer; the inhibition of the transfer is comparison to the controlbatch without substance is determined

B-I.3. Obtainment of Radiolabelled HDL

50 ml of fresh human EDTA plasma is adjusted to a density of 1.12 usingNaBr and centrifuged at 4° C. in a Ty 65 rotor at 50 000 rpm for 18 h.The upper phase is used for the obtainment of cold LDL. The lower phaseis dialysed against 3×41 of PDB buffer (10 mM tris/HCl pH 7.4, 0.15 mMNaCl, 1 mM EDTA, 0.02% NaN₃). Per 10 ml of retentate volume, 20 μl of³H-cholesterol (Dupont NET-725; 1 μC/μl dissolved in ethanol) are thenadded and the mixture is incubated at 37° C. under N₂ for 72 h.

The batch is then adjusted to the density 1.21 using NaBr andcentrifuged at 20° C. in a Ty 65 rotor at 50 000 rpm for 18 h. The upperphase is recovered and the lipoprotein fractions are purified bygradient centrifugation. To this end, the isolated, labelled lipoproteinfraction is adjusted to a density of 1.26 using NaBr. 4 ml each of thissolution are covered in centrifuge tubes (SW 40 rotor) with 4 ml of asolution of density 1.21 and 4.5 ml of a solution of density 1.063(density solutions of PDB buffer and NaBr) and then centrifuged for 24 hat 38 000 rpm and 20° C. in the SW 40 rotor. The intermediate layerlying between the density 1.063 and 1.21, containing the labelled HDL,is dialysed against 3×100 volumes of PDB buffer at 4° C.

The retentate contains radiolabelled ³H-CE-HDL, which, adjusted to about5×10⁶ cmp per ml, is used for the test.

B-I.4. CETP-SPA Test

For testing of the CETP activity, the transfer of ³H-cholesterol esterfrom human HD lipoproteins to biotinylated LD lipoproteins is measured.The reaction is ended by addition of streptavidin-SPA® beads (Amersham)and the transferred radioactivity is determined directly in a liquidscintillation counter.

In the test batch, 10 μl of HDL-³H-cholesterol ester (˜50 000 cpm) areincubated at 37° C. for 18 h with 10 μl of biotin-LDL (Amersham) in 50mM Hepes/0.15 M NaCl/0.1% bovine serum albumin/0.05% NaN₃ pH 7.4containing 10 μl of CETP (1 mg/ml) and 3 μl of a solution of thesubstance to be tested (dissolved in 10% DMSO/1% RSA). 200 μl of theSPA-streptavidin bead solution (TRKQ 7005) are then added, incubatedfurther with shaking for 1 h and then measured in a scintillationcounter. Corresponding incubations with 10 μl of buffer, 10 μl of CETPat 4° C. and 10 μl of CETP at 37° C. serve as controls.

The activity transferred in the control batches with CETP at 37° C. israted as 100% transfer. The substance concentration at which thistransfer is reduced to half is specified as the IC₅₀ value.

Example IC₅₀ [nM] No. SPA Test 1 7B-II.1. Measurement of the Ex Vivo Activities on Transgenic hCETP Mice

To test for CETP-inhibitory activity, the substances are administeredorally using a stomach tube to transgenic hCETP mice bred in-house[Dinchuk et al. BBA 1295-301 (1995)]. To this end, male animals arerandomly assigned to groups having an equal number of animals, as a rulen=4, one day before the start of the experiment. Before administrationof the substance, blood is taken from each mouse by puncture of theretro-orbital venous plexus for the determination of its basal CETPactivity in the serum (T1). The test substance is then administered tothe animals using the stomach tube. At specific times afteradministration of the test substance, blood is taken from the animals bypuncture a second time (T2), in general 16 or 24 h after substanceadministration, but if appropriate this can also be carried out atanother time.

In order to be able to assess the inhibitory activity of a substance,for each time, i.e. 16 or 24 hours, a corresponding control group isemployed whose animals only receive the formulating agent withoutsubstance. In the control animals, the second blood sampling per animalis carried out as in the substance-treated animals in order to be ableto determine the change in the CETP activity without inhibitor over thecorresponding experimental time interval (16 or 24 h).

After termination of the clotting, the blood samples are centrifuged andthe serum is removed by pipette. For the determination of the CETPactivity, the cholesteryl ester transport over 4 h to is determined. Tothis end, in general 2 μl of serum are employed in the test batch andthe test is carried out as described under B-I.2.3.

The differences in the cholesteryl ester transport [pM CE/h (T2)−pM CE/h(T1)] are calculated for each animal and averaged in the groups. Asubstance which at one of the times reduces the cholesteryl estertransport by >20% is regarded as active.

% inhibition Example at 3 mg/kg No. 16 h 24 h 1 50 24

B-II.2. Measurement of the In Vivo Activity in Syrian Golden Hamsters

Female Syrian golden hamsters bred in-house (strain BAY:DSN) and havinga weight of 150-200 g are used to determine the oral action of CETPinhibitors on serum lipoproteins and triglycerides. The animals aregrouped in six animals per cage and acclimatized to feed and water adlibitum for two weeks.

Immediately prior to the start of the experiment and after the substancehas been administered, blood is withdrawn by retro-orbital puncture ofthe venous plexus and used to obtain serum after 30 min of incubation atroom temperature and 20 min of centrifugation at 30 000 g. Thesubstances are dissolved in 20% Solutol/80% water and administered perorally by means of a stomach tube. The controlled animals receiveidentical volumes of solvent without test substance.

Triglycerides, total cholesterol, HDL cholesterol and LDL cholesterolare determined using the analytical instrument COBAS INTEGRA 400 plus(from Roche Diagnostics) according to the instructions of themanufacturer. From the measured values, for each parameter, the changein percent caused by the treatment with the substance is calculated foreach animal and stated as mean with standard deviation per group (n=6 orn=12). If, compared to the group treated with solvent, the effects ofthe substance are significant, the p-value to determined by applicationof the t-test is added (* p ≦0.05; ** p ≦0.01; *** p ≦0.005).

Example % increase of HDL after No. 24 h (dose: 2 × 10 mg/kg) 1 23B-II.3. Measurement of the In Vivo Activity in Transgenic hCETP Mice

To determine the oral action on lipoproteins and triglycerides, testsubstance is administered to transgenic mice [Dinchuk et al., BBA,1295-1301 (1995)] using a stomach tube. Before the start of theexperiment, blood is withdrawn from the mice retro-orbitally in order todetermine cholesterol and triglycerides in the serum. The serum isobtained as described above for hamsters by incubation at 4° C.overnight and subsequent centrifugation at 6000 g. After three days,blood is again withdrawn from the mice in order to determinelipoproteins and triglycerides. The changes in the parameters measuredare expressed as the percentage change compared with the starting value.

Example % increase of HDL after No. 3 d (dose: 3 × 3 mg/kg) 1 91

C. WORKING EXAMPLES OF PHARMACEUTICAL COMPOSITIONS (a)

The compound of the invention can be converted into pharmaceuticalpreparations in the following ways:

Tablet: Composition:

100 mg of the compound of the invention, 50 mg of lactose (monohydrate),50 mg of maize starch (native), 10 mg of polyvinylpyrrolidone (PVP 25)(from BASF, Ludwigshafen, Germany) and 2 mg of magnesium stearate.

Tablet weight 212 mg, diameter 8 mm, radius of curvature 12 mm.

Production:

The mixture of compound of the invention, lactose and starch isgranulated with a 5% strength solution (m/m) of the PVP in water. Thegranules are dried and mixed with the magnesium stearate for 5 minutes.This mixture is compressed in a conventional tablet press (see above forformat of the tablet). A guideline compressive force for the compressionis 15 kN.

Suspension which can be Administered Orally:

Composition:

1000 mg of the compound of the invention, 1000 mg of ethanol (96%), 400mg of Rhodigel® (xanthan gum from FMC, Pennsylvania, USA) and 99 g ofwater.

10 ml of oral suspension correspond to a single dose of 100 mg of thecompound of the invention.

Production:

The Rhodigel is suspended in ethanol, and the compound of the inventionis added to the suspension. The water is added while stirring. Themixture is stirred for about 6 h until the swelling of the Rhodigel iscomplete.

Solution which can be Administered Orally:

Composition:

500 mg of the compound of the invention, 2.5 g of polysorbate and 97 gof polyethylene glycol 400.20 g of oral solution correspond to a singledose of 100 mg of the compound of the invention.

Production:

The compound of the invention is suspended in the mixture ofpolyethylene glycol and polysorbate with stirring. The stiffing processis continued until the compound of the invention has completelydissolved.

i.v. Solution:

The compound of the invention is dissolved in a concentration below thesaturation solubility in a physiologically tolerated solvent (e.g.isotonic saline, 5% glucose solution and/or 30% PEG 400 solution). Thesolution is sterilized by filtration and used to fill sterile andpyrogen-free injection containers.

Starting Materials and Intermediates: (b) Example 1A4-cyclopentyl-2-isopropyl-7,7-dimethyl-3-[4-(trifluoromethyl)benzoyl]-4,6,7,8-tetrahydroquinolin-5(1H)-one

10.0 g (38.9 mmol, 1.0 eq.) of3-amino-3-isopropyl-1-(4-trifluoromethylphenyl)propenone (preparationaccording to WO 03/028727, Example 2) are initially charged in 300 ml ofdiisopropyl ether, and 2.99 ml (38.9 mmol, 1.0 eq.) of trifluoroaceticacid and 5.45 g (38.9 mmol, 1 eq.) of 5,5-dimethylcyclohexane-1,3-dioneare added. After 10 min of stirring at room temperature, the mixture isheated to reflux and 4.58 g (46.7 mmol, 1.2 eq.) ofcyclopentanecarbaldehyde are added. The mixture is heated under refluxon a water separator for 15 h. After cooling, the mixture is stirred inan ice bath for 45 min and the resulting precipitate is filtered offwith suction, washed with cold diisopropyl ether and freed from solventresidues under high vacuum.

Yield: 4.15 g (23% of theory)

¹H-NMR (CDCl₃, 400 MHz): δ=1.05 (d, 3H), 1.15 (s, 3H), 1.16 (s, 3H),1.28 (d, 3H), 1.24-1.61 (m, 7H), 2.30 and 2.51 (2d, 2H), 2.34 (s, 2H),3.49 (sept, 1H), 3.81 (d, 1H), 5.96 (s, 1H), 7.66 (d, 2H), 7.77 (d, 2H)ppm.

MS (DCI/NH₃): m/z=460 [M+H]⁺, 477 [M+NH₄]⁺.

Example 2A4-cyclopentyl-2-isopropyl-7,7-dimethyl-3-[4-(trifluoromethyl)benzoyl]-7,8-dihydroquinolin-5(6H)-one

4.0 g (8.7 mmol) of the compound from Example 1A are dissolved in 100 mlof dichloro-methane, and 2.17 g (9.6 mmol, 1.1 eq.) of2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) are added a little at atime at 0° C. With stirring, the mixture is warmed to room temperatureover a period of 3 h. The mixture is concentrated on a rotary evaporatorand the residue is purified by chromatography (silica gel, mobilephase:isohexane/ethyl acetate 100:0→50:50).

Yield: 3.78 g (95.1% of theory)

¹H-NMR (CDCl₃, 300 MHz): δ=1.09 (d, 3H), 1.11 (s, 3H), 1.17 (s, 3H),1.19 (d, 3H), 1.34-2.00 (m, 8H), 2.51-2.68 (m, 3H), 3.01 (m, 1H), 3.1(s, 2H), 7.76 (d, 2H), 7.94 (m, 2H) ppm.

MS (ESIpos): m/z=458 [M+H]⁺.

Example 3A[(5S)-4-cyclopentyl-5-hydroxy-2-isopropyl-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-3-yl][4-(trifluoromethyl)phenyl]methanone

140 mg (0.96 mmol, 0.08 eq.) of (1R,2S)-1-aminoindan-2-ol are initiallycharged in 440 ml of THF, and 7.80 g (47.8 mmol, 4.0 eq.) of boraneN,N-diethylaniline complex are added at room temperature. After theevolution of gas has ceased, the mixture is cooled to 0° C. and 5.47 g(12 mmol, 1 eq.) of the compound from Example 2A, dissolved in 40 ml ofTHF, are added. With stirring, the mixture is allowed to warm to roomtemperature over a period of 28 h. After the reaction has ended, 20 mlof methanol are added to the reaction mixture and the mixture isconcentrated. The residue is partitioned between 150 ml of water and 150ml of ethyl acetate. The aqueous phase is extracted twice with in eachcase 100 ml of ethyl acetate. The combined organic phases are washedwith 50 ml of saturated sodium chloride solution, dried over sodiumsulphate, filtered and concentrated under reduced pressure. The crudeproduct is then purified by chromatography (silica gel, mobilephase:isohexane/ethyl acetate 4:1).

Yield: 4.97 g (90.5% of theory)

The enantiomeric excess is determined according to method 1 as being92.5% ee.

The enantiomers are separated by chromatography on a chiral phase(column: Chiralpak AD, 500 mm×40 mm, 20 μm; mobilephase:isohexane/isopropanol 97.5:2.5; flow rate: 50 ml/min):

Yield: 4.46 g (81.2% of theory)

The enantiomeric excess is determined according to method 1 as being98.1% ee; R_(t) (method 1)=7.09 min.

¹H-NMR (CDCl₃, 300 MHz): δ=0.94-1.30 (m, 12H), 1.31-2.03 (m, 11H), 2.53(m, 1H), 2.72 (d, 1H), 2.95-3.12 (m, 1H), 3.29 (m, 1H), 5.18 (m, 1H),7.73 (d, 2H), 7.93 (m, 2H) ppm.

MS (DCI): m/z=460 [M+H]⁺.

Example 4A((5S)-5-{[tert-butyl(dimethyl)silyl]oxy}-4-cyclopentyl-2-isopropyl-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-3-yl)[4-(trifluoromethyl)phenyl]methanone

Under argon, 3.65 g (7.95 mmol) of the compound from Example 3A areinitially charged in 80 ml of dry toluene. At room temperature, 3.41 g(31.8 mmol, 4 eq.) of 2,6-lutidine are then added, and the mixture iscooled to −18° C. 3.65 ml (15.9 mmol, 2 eq.) of tert-butyldimethylsilyltrifluoromethanesulphonate are added dropwise to this solution. After 20min, the reaction mixture is warmed to 0° C. and stirred at thistemperature for a further 55 min. For work-up, saturated ammoniumchloride solution (100 ml) is added and the mixture is, after warming toroom temperature, extracted with ethyl acetate. The aqueous phase isextracted two more times with ethyl acetate and the combined organicphases are washed with saturated sodium chloride solution, dried oversodium sulphate, filtered and concentrated under reduced pressure. Theresidue is purified by chromatography (silica gel, mobilephase:isohexane/ethyl acetate 9:1).

Yield: 4.65 g (quantitative)

¹H-NMR (CDCl₃, 400 MHz): δ=0.09 (s, 3H), 0.18 (s, 3H), 0.86 (s, 9H),0.92 (s, 3H), 1.04 (d, 3H), 1.22 (d, 3H), 1.24 (s, 3H), 1.27-1.86 (m,9H), 1.93-2.02 (m, 1H), 2.50 (m, 1H), 2.58-3.23 (m, 2H), 3.32 (m, 1H),5.20 (m, 1H), 7.73 (d, 2H), 7.83-8.00 (m, 2H) ppm.

MS (ESIpos): m/z=574 [M+H]⁺.

Example 5A(S)-((5S)-5-{[tert-butyl(dimethyl)silyl]oxy}-4-cyclopentyl-2-isopropyl-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-3-yl)[4-(trifluoromethyl)phenyl]methanol

At 0° C., 12.0 ml of a 1 M solution of lithium aluminium hydride (12.0mmol, 1.5 eq.) in THF are added dropwise to a solution of 4.59 g (8.0mmol) of the compound from Example 4A in 80 ml of dry THF. Withstirring, the mixture is warmed to room temperature over a period of 16h. For work-up, 120 ml of a saturated sodium potassium tartrate solutionare added carefully. After the evolution of gas has ceased, the mixtureis extracted twice with ethyl acetate and the combined organic phasesare washed with saturated sodium chloride solution, dried over sodiumsulphate, filtered and concentrated under reduced pressure. The residueis purified chromatographically and at the same time separated into thediastereomers of the product (silica gel, mobile phase:isohexane/ethylacetate 95:5).

Yield: 2.29 g (49.8% of theory)

¹H-NMR (CDCl₃, 300 MHz): δ=0.13 (s, 3H), 0.20 (s, 3H), 0.64-1.00 (m,18H), 1.09-2.23 (m, 14H), 2.59 (d, 1H), 2.89 (m, 1H), 3.00 (m, 1H), 3.71(m, 1H), 5.21 (t, 1H), 6.22 (br. s, 1H), 7.43 (d, 2H), 7.59 (d, 2H) ppm.

MS (ESIpos): m/z=576 [M+H]⁺

R_(f)=0.26 (isohexane/ethyl acetate 9:1).

Syn Diastereomer(R)-((5S)-5-{[tert-butyl(dimethyl)silyl]oxy}-4-cyclopentyl-2-isopropyl-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-3-yl)[4-(trifluoromethyl)phenyl]methanol

Yield: 2.48 g (53.9% of theory)

R_(f)=0.34 (isohexane/ethyl acetate 9:1).

Example 6A(5S)-5-{[tert-butyl(dimethyl)silyl]oxy}-4-cyclopentyl-3-{(S)-fluoro[4-(trifluoromethyl)phenyl]methyl}-2-isopropyl-7,7-dimethyl-5,6,7,8-tetrahydroquinoline

At −10° C. and under argon, 0.82 ml of diethylaminosulphur trifluoride(6.2 mmol, 1.5 eq.) is added dropwise to a solution of 2.38 g (4.1 mmol)of the compound from Example 5A in 40 ml of dry dichloromethane. Themixture is stirred at this temperature for 200 min. For work-up, 40 mlof a saturated sodium bicarbonate solution are carefully added with icecooling. The mixture is extracted three times in total with ethylacetate. The combined organic phases are then washed with saturatedsodium chloride solution, dried over sodium sulphate, filtered andconcentrated under reduced pressure. The crude product is purified byfiltration through silica gel (mobile phase:cyclohexane/ethyl acetate9:1).

Yield: 1.92 g (80.3% of theory)

LC/MS (method 3): R_(t)=3.85 min.

MS (ESIpos): m/z=578 [M+H]⁺

R_(f)=0.66 (isohexane/ethyl acetate 9:1).

WORKING EXAMPLES: (b) Example 1(5S)-4-cyclopentyl-3-{(S)-fluoro[4-(trifluoromethyl)phenyl]methyl}-2-isopropyl-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-5-ol

At 0° C., 13.3 ml of a 1 M solution of tetrabutylammonium fluoride (13.3mmol, 4.0 eq.) in THF are added dropwise to a solution of 1.92 g (3.3mmol) of the compound from Example 6A in 20 ml of dry THF. The reactionmixture is stirred in an ice bath for 4 h. For work-up, the mixture isdiluted with 100 ml of ethyl acetate and washed twice with in each case100 ml of water and with 50 ml of saturated sodium chloride solution.The organic phase is dried over sodium sulphate, filtered andconcentrated under reduced pressure. The crude product is purifiedchromatographically (silica gel, mobile phase:isohexane/ethyl acetate9:1→2:1).

Yield: 1.18 g (76.4% of theory)

¹H-NMR (CDCl₃, 300 MHz): δ=0.70 (d, 3H), 1.03 (s, 3H), 1.13 (d, 3H),1.19 (s, 3H), 1.32-2.20 (m, 11H), 2.63-2.97 (m, 3H), 3.86 (m, 1H), 5.15(m, 1H), 6.94 (d, 1H), 7.34 (d, 2H), 7.61 (d, 2H) ppm.

MS (DCI): m/z=464 [M+H]⁺

R_(f)=0.13 (isohexane/ethyl acetate 4:1).

The further separation of diastereomers still present in the product iscarried out by chromatography (column: Chiralpak AD, 500 mm×40 mm, 20μm; mobile phase:isohexane/ethyl 97.5:2.5; flow rate: 50 ml/min).

Yield: 0.35 g (22.9% of theory).

B. ASSESSMENT OF THE PHARMACOLOGICAL ACTIVITY (b) B-I. CETP-InhibitionTesting B-I.1. Obtainment of CETP

CETP is obtained in partially purified form from human plasma bydifferential centrifugation and column chromatography and used for thetest. To this end, human plasma is adjusted to a density of 1.21 g perml using NaBr and centrifuged at 4° C. at 50 000 rpm for 18 h. Thebottom fraction (d >1.21 g/ml) is applied to aSephadex®-Phenyl-Sepharose 4B (Pharmacia) column, washed with 0.15 MNaCl/0.001 M tris/HCl pH 7.4 and then eluted with distilled water. TheCETP-active fractions are pooled, dialysed against 50 mM sodium acetatepH 4.5 and applied to a CM-Sepharose® column (Pharmacia). The mixture isthen eluted using a linear gradient (0-1 M NaCl). The pooled CETPfractions are dialysed against 10 mM tris/HCl pH 7.4 and then furtherpurified by chromatography on a Mono Q® column (Pharmacia).

B-I.2. CETP Fluorescence Test

Measurement of the CETP-catalysed transfer of a fluorescent cholesterolester between liposomes [modified according to the procedure of Bisgaieret al., J. Lipid Res. 34, 1625 (1993)]:

For the production of the donor liposomes, 1 mg of cholesteryl4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-dodecanoate(cholesteryl BODIPY® FL C₁₂, Molecular Probes) is dissolved in 600 μl ofdioxane with 5.35 mg of triolein and 6.67 mg of phosphatidylcholine withgentle warming in an ultrasonic bath and this solution is added veryslowly with ultrasonication to 63 ml of 50 mM tris/HCl, 150 mM NaCl, 2mM EDTA buffer pH 7.3 at room temperature. The suspension is thensonicated under an N₂ atmosphere for 30 minutes in the Bransonultrasonic bath at about 50 watts, the temperature being kept at about20° C.

The acceptor liposomes are obtained analogously from 86 mg ofcholesteryl oleate, 20 mg of triolein and 100 mg of phosphatidylcholinedissolved in 1.2 ml of dioxane and 114 ml of the above buffer byultrasonication at 50 watts (20° C.) for 30 minutes.

B-I.2.1. CETP Fluorescence Test with Enriched CETP

For testing, a test mix consisting of 1 part of above buffer, 1 part ofdonor liposomes and 2 parts of acceptor liposomes is used.

50 μl of test mix are treated with 48 μl of enriched CETP fraction (1-3μg), obtained from human plasma by means of hydrophobic chromatography,and 2 μl of a solution of the substance to be investigated in DMSO andincubated at 37° C. for 4 hours.

The change in the fluorescence at 485/535 nm is a measure of the CEtransfer; the inhibition of the transfer in comparison to the controlbatch without substance is determined

IC₅₀ [nM] Example fluorescence No. test 1 25B-I.2.2. CETP Fluorescence Test with Human Plasma

6 μl (12% v/v) of donor liposomes and 1 μl (2% v/v) of a solution of thesubstance to be investigated in DMSO are added to 42 μl (86% v/v) ofhuman plasma (Sigma P9523), and the mixture is incubated at 37° C. for24 h.

The change in the fluorescence at 510/520 nm (gap width 2.5 nm) is ameasure of the CE transfer; the inhibition of the transfer in comparisonto the control batch without substance is determined

IC₅₀ [nM] Example fluorescence test in No. human plasma 1 84

B-I.2.3. Ex Vivo-CETP Fluorescence Test

10 μl of buffer and 2 μl of serum are added to 80 μl of test mix, andthe mixture is incubated at 37° C. for 4 h.

The change in the fluorescence at 485/535 nm is a measure for the CEtransfer; the inhibition of the transfer in comparison to the controlbatch without substance is determined.

B-I.3. Obtainment of Radiolabelled HDL

50 ml of fresh human EDTA plasma are adjusted to a density of 1.12 usingNaBr and centrifuged at 4° C. in a Ty 65 rotor at 50 000 rpm for 18 h.The upper phase is used for the obtainment of cold LDL. The lower phaseis dialysed against 3×41 of PDB buffer (10 mM tris/HCl pH 7.4, 0.15 mMNaCl, 1 mM EDTA, 0.02% NaN₃). Per 10 ml of retentate volume, 20 μl of³H-cholesterol (Dupont NET-725; 1 μC/μl dissolved in ethanol) are thenadded and the mixture is incubated at 37° C. under N₂ for 72 h.

The batch is then adjusted to the density 1.21 using NaBr andcentrifuged at 20° C. in a Ty 65 rotor at 50 000 rpm for 18 h. The upperphase is recovered and the lipoprotein fractions are purified bygradient centrifugation. To this end, the isolated, labelled lipoproteinfraction is adjusted to a density of 1.26 using NaBr. 4 ml each of thissolution are covered in centrifuge tubes (SW 40 rotor) with 4 ml of asolution of density 1.21 and 4.5 ml of a solution of density 1.063(density solutions of PDB buffer and NaBr) and then centrifuged for 24 hat 38 000 rpm and 20° C. in the SW 40 rotor. The intermediate layerlying between the density 1.063 and 1.21, containing the labelled HDL,is dialysed against 3×100 volumes of PDB buffer at 4° C.

The retentate contains radiolabelled ³H-CE-HDL, which, adjusted to about5×10⁶ cmp per ml, is used for the test.

B-I.4. CETP-SPA Test

For testing of the CETP activity, the transfer of ³H-cholesterol esterfrom human HD lipoproteins to biotinylated LD lipoproteins is measured.The reaction is ended by addition of streptavidin-SPA® beads (Amersham)and the transferred radioactivity is determined directly in a liquidscintillation counter.

In the test batch, 10 μl of HDL-³H-cholesterol ester (˜50 000 cpm) areincubated at 37° C. for 18 h with 10 μl of biotin-LDL (Amersham) in 50mM Hepes/0.15 M NaCl/0.1% bovine serum albumin/0.05% NaN₃ pH 7.4containing 10 μl of CETP (1 mg/ml) and 3 μl of a solution of thesubstance to be tested (dissolved in 10% DMSO/1% RSA). 200 μl of theSPA-streptavidin bead solution (TRKQ 7005) are then added, incubatedfurther with shaking for 1 h and then measured in a scintillationcounter. Corresponding incubations with 10 μl of buffer, 10 μl of CETPat 4° C. and 10 μl of CETP at 37° C. serve as controls.

The activity transferred in the control batches with CETP at 37° C. israted as 100% transfer. The substance concentration at which thistransfer is reduced to half is specified as the IC₅₀ value.

Example IC₅₀ [nM] No. SPA Test 1 12B-II.1. Measurement of the Ex Vivo Activities on Transgenic hCETP Mice

To test for CETP-inhibitory activity, the substances are administeredorally using a stomach tube to transgenic hCETP mice bred in-house[Dinchuk et al., BBA, 1295-1301 (1995)]. To this end, male animals arerandomly assigned to groups having an equal number of animals, as a rulen=4, one day before the start of the experiment. Before administrationof the substance, blood is taken from each mouse by puncture of theretro-orbital venous plexus for the determination of its basal CETPactivity in the serum (T1). The test substance is then administered tothe animals using the stomach tube. At specific times afteradministration of the test substance, blood is taken from the animals bypuncture a second time (T2), in general 16 or 24 h after substanceadministration, but if appropriate this can also be carried out atanother time.

In order to be able to assess the inhibitory activity of a substance,for each time, i.e. 16 or 24 hours, a corresponding control group isemployed whose animals only receive the formulating agent withoutsubstance. In the control animals, the second blood sampling per animalis carried out as in the substance-treated animals in order to be ableto determine the change in the CETP activity without inhibitor over thecorresponding experimental time interval (16 or 24 h).

After termination of the clotting, the blood samples are centrifuged andthe serum is removed by pipette. For the determination of the CETPactivity, the cholesteryl ester transport over 4 h is determined. Tothis end, in general 2 μl of serum are employed in the test batch andthe test is carried out as described under B-I.2.3.

The differences in the cholesteryl ester transport [pM CE/h (T2)−pM CE/h(T1)] are calculated for each animal and averaged in the groups. Asubstance which at one of the times reduces the cholesteryl estertransport by >20% is regarded as active.

% inhibition Example at 3 mg/kg No. 16 h 24 h 1 49 39

B-II.2. Measurement of the In Vivo Activity in Syrian Golden Hamsters

Female Syrian golden hamsters bred in-house (strain BAY:DSN) and havinga weight of 150-200 g are used to determine the oral action of CETPinhibitors on serum lipoproteins and triglycerides. The animals aregrouped in six animals per cage and acclimatized to feed and water adlibitum for two weeks.

Immediately prior to the start of the experiment and after the substancehas been administered, blood is withdrawn by retro-orbital puncture ofthe venous plexus and used to obtain serum after 30 min of incubation atroom temperature and 20 min of centrifugation at 30 000 g. Thesubstances are dissolved in 20% Solutol/80% water and administeredperorally by means of a stomach tube. The control animals receiveidentical volumes of solvent without test substance.

Triglycerides, total cholesterol, HDL cholesterol and LDL cholesterolare determined using the analytical instrument COBAS INTEGRA 400 plus(from Roche Diagnostics) according to the instructions of themanufacturer. From the measured values, for each parameter, the changein percent caused by the treatment with the substance is calculated foreach animal and stated as mean with standard deviation per group (n=6 orn=12). If, compared to the group treated with solvent, the effects ofthe substance are significant, the p-value determined by application ofthe t-test is added (* p ≦0.05; ** p ≦0.01; *** p ≦0.005).

Example % increase of HDL after No. 24 h (dose: 2 × 10 mg/kg) 1 20B-II.3. Measurement of the In Vivo Activity in Transgenic hCETP Mice

To determine the oral action on lipoproteins and triglycerides, testsubstance is administered to transgenic mice [Dinchuk et al., BBA,1295-1301 (1995)] using a stomach tube. Before the start of theexperiment, blood is withdrawn from the mice retro-orbitally in order todetermine cholesterol and triglycerides in the serum. The serum isobtained as described above for hamsters by incubation at 4° C.overnight and subsequent centrifugation at 6000 g. After three days,blood is again withdrawn from the mice in order to determinelipoproteins and triglycerides. The changes in the parameters measuredare expressed as the percentage change compared with the starting value.

Example % increase of HDL after No. 3 d (dose: 3 × 3 mg/kg) 1 85

C. WORKING EXAMPLES OF PHARMACEUTICAL COMPOSITIONS (b)

The compounds of the invention can be converted into pharmaceuticalpreparations in the following ways:

Tablet: Composition:

100 mg of the compound of the invention, 50 mg of lactose (monohydrate),50 mg of maize starch (native), 10 mg of polyvinylpyrrolidone (PVP 25)(from BASF, Ludwigshafen, Germany) and 2 mg of magnesium stearate.

Tablet weight 212 mg, diameter 8 mm, radius of curvature 12 mm

Production:

The mixture of compound of the invention, lactose and starch isgranulated with a 5% strength solution (m/m) of the PVP in water. Thegranules are dried and mixed with the magnesium stearate for 5 minutes.This mixture is compressed in a conventional tablet press (see above forformat of the tablet). A guideline compressive force for the compressionis 15 kN.

Suspension which can be Administered Orally:

Composition:

1000 mg of the compound of the invention, 1000 mg of ethanol (96%), 400mg of Rhodigel® (xanthan gum from FMC, Pennsylvania, USA) and 99 g ofwater.

10 ml of oral suspension correspond to a single dose of 100 mg of thecompound of the invention.

Production:

The Rhodigel is suspended in ethanol, and the compound of the inventionis added to the suspension. The water is added while stirring. Themixture is stirred for about 6 h until the swelling of the Rhodigel iscomplete.

Solution which can be Administered Orally:

Composition:

500 mg of the compound of the invention, 2.5 g of polysorbate and 97 gof polyethylene glycol 400.20 g of oral solution correspond to a singledose of 100 mg of the compound of the invention.

Production:

The compound of the invention is suspended in the mixture ofpolyethylene glycol and polysorbate with stiffing. The stirring processis continued until the compound of the invention has completelydissolved.

i.v. Solution:

The compound of the invention is dissolved in a concentration below thesaturation solubility in a physiologically tolerated solvent (e.g.isotonic saline, 5% glucose solution and/or 30% PEG 400 solution). Thesolution is sterilized by filtration and used to fill sterile andpyrogen-free injection containers.

1-10. (canceled)
 11. A method for the primary and/or secondaryprevention of coronary heart disease in a humans or animal comprisingadministering an effective amount of a compound of formula (I)

in which R represents cyclopentyl or isopropyl, and salts thereof.
 12. Amethod for the treatment and/or prevention of hypolipoproteinaemias,dyslipidaemias, hypetriglyceridaemias, hyperlipidaemias,hypercholesterolaemias, arteriosclerosis, restenosis, adiposity,obesity, diabetes, stroke and Alzheimer's disease in a humans or animalby administering an effective amount of a compound of formula (I).

in which R represents cyclopentyl or isopropyl, and salts thereof.13-14. (canceled)
 15. The method of claim 11, wherein the compound offormula (I) is administered as a pharmaceutical composition comprisingthe compound in combination with an inert nontoxic pharmaceuticallysuitable auxiliary.
 16. The method of claim 11, wherein the compound offormula (I) is administered as a pharmaceutical composition comprisingthe compound and at least one active compound selected from the groupconsisting of an antidiabetic, a platelet aggregation inhibitor, ananticoagulant, a calcium antagonist, an angiotensin AII antagonist, anACE inhibitor, a beta blocker, a phosphodiesterase inhibitor, astimulator of soluble guanylate cyclase, a cGMP enhancer, a diuretic, athyroid receptor agonist, an HMG-CoA reductase inhibitor, a squalenesynthase inhibitor, a squalene epoxidase inhibitor, a, oxidosqualenecyclase inhibitor, an ACAT inhibitor, a MTP inhibitor, a PPAR agonist, afibrate, a lipase inhibitor, a cholesterol absorption inhibitor, a bileacid reabsorption inhibitor, a polymeric bile acid adsorber and alipoprotein(a) antagonist.
 17. The method of claim 12, wherein thecompound of formula (I) is administered as a pharmaceutical compositioncomprising the compound in combination with an inert nontoxicpharmaceutically suitable auxiliary.
 18. The method of claim 12, whereinthe compound of formula (I) is administered as a pharmaceuticalcomposition comprising the compound and at least one active compoundselected from the group consisting of an antidiabetic, a plateletaggregation inhibitor, an anticoagulant, a calcium antagonist, anangiotensin AII antagonist, an ACE inhibitor, a beta blocker, aphosphodiesterase inhibitor, a stimulator of soluble guanylate cyclase,a cGMP enhancer, a diuretic, a thyroid receptor agonist, an HMG-CoAreductase inhibitor, a squalene synthase inhibitor, a squalene epoxidaseinhibitor, a, oxidosqualene cyclase inhibitor, an ACAT inhibitor, a MTPinhibitor, a PPAR agonist, a fibrate, a lipase inhibitor, a cholesterolabsorption inhibitor, a bile acid reabsorption inhibitor, a polymericbile acid adsorber and a lipoprotein(a) antagonist.